有關(guān)我院醫(yī)學本科生基礎學習模塊生物化學部分相關(guān)教師

1、有關(guān)我院醫(yī)學本科生基礎學習模塊生物化學部分相有關(guān)我院醫(yī)學本科生基礎學習模塊生物化學部分相關(guān)教師授課期間考察學生的決定以及實施參考意見關(guān)教師授課期間考察學生的決定以及實施參考意見 為了教師在授課期間，加強對教學過程的有效管理與靈為了教師在授課期間，加強對教學過程的有效管理與靈活操作，充分調(diào)動學生學習生物化學課程的積極性與主動性，活操作，充分調(diào)動學生學習生物化學課程的積極性與主動性，經(jīng)征求科教處同意，決定在生物化學授課期間由授課教師對經(jīng)征求科教處同意，決定在生物化學授課期間由授課教師對學生的學習情況實施考察，并對每一個聽課學生記錄一個成學生的學習情況實施考察，并對每一個聽課學生記錄一個成績，并將該

2、成績最終記入期末總成績中?？?，并將該成績最終記入期末總成績中。無故不來上課者扣分。無故不來上課者扣分。上課時，未經(jīng)請假提前早退者扣分。上課時，未經(jīng)請假提前早退者扣分。上課時，影響教師授課氛圍者扣分。上課時，影響教師授課氛圍者扣分。上課遲到者扣分。上課遲到者扣分。對上課討論時主動發(fā)言，積極提問或出色完成授課教對上課討論時主動發(fā)言，積極提問或出色完成授課教師所布置的作業(yè)或其它相關(guān)學習任務，且無上述不良師所布置的作業(yè)或其它相關(guān)學習任務，且無上述不良情況者情況者,每位教師根據(jù)具體情況可給予這樣的學生鼓勵每位教師根據(jù)具體情況可給予這樣的學生鼓勵加分并記入總成績，但最終總成績以加分并記入總成績，但最終總成

3、績以100分為限。分為限。 為了此項考查的客觀公證性，使該舉措真正發(fā)揮作用。特為了此項考查的客觀公證性，使該舉措真正發(fā)揮作用。特提出如下操作規(guī)則提出如下操作規(guī)則10. DNA replication*11. RNA transcription*12. Protein biosynthesis*13. Regulation of gene expression*14. Gene recombination and gene engineeringDNAs RNAs Proteins10 DNA replication: entirety11 RNA transcription: systemat

4、icness12 Protein biosynthesis: individuality 10101211Chapter tenDNA replication Section 1* General features of DNA replicationSection 2* Enzymes in DNA replication Section 3 Process of DNA replicationSection 4 Reverse transcription and othersSection 5* DNA damage and repairConcept of DNA replication

5、 or DNA biosynthesis The transmission of the genetic informations between DNA and DNA The synthesis of a complementary DNA strand by forming phosphodiester linkages between nucleotides base-paired to template strand is catalyzed by large multienzyme complexes referred to as the DNA polymerases Semic

6、onservative replication Section 1General features of DNA replication1) Experimental basis and the significance of semiconservative replication*2) DNA replication is Bidirectional 3) Semidiscontinuous pattern of DNA replication 1) Experimental basis and the significance of semiconservative replicatio

7、nA celebrated scientific experiment daughter DNAmoleculesparent DNAmoleculesparent DNAstrandDaughter or new DNAstrand total conservesemi conserve commingle The basic experiment condition A experimental table A centrifuge A kind of bacterium A few 14NH4Cl culture solutions A few 15NH4Cl culture solut

8、ions A few sucroses7N14,15 / 1S22S22P3The experiment procedurebacterium15cultureextraction bacteriumDNAcentrifugationDNADNADNAbacteriumbacteriumDNA layerDNA layerDNA layerDNA layerextraction extraction extraction centrifugationcentrifugationcentrifugation14Nculture14Nculture14N1234total conserveLL+H

9、1515HLLLL+HL+HL+H14141414141514141414141415LL14/1514/1514/151514/1514/1514/15LMM+LM+LM+LHM+Lsemi conserve1414141414141414LLThe experiment procedurebacterium14cultureextraction bacteriumDNAcentrifugationDNADNADNAbacteriumbacteriumDNA layerDNA layerDNA layerDNA layerextraction extraction extraction ce

10、ntrifugationcentrifugationcentrifugation15Nculture15Nculture15N123414/1514/1514151514/1515151514/151514/15151514/15HHLMM+HM+HM+HHM+Hsemi conserve Significance of semiconservative replication Genetic conservativenessACGTTGCAACGTTGCAACGTTGCASection 2Enzymes in DNA replicationChemical reaction in DNA r

11、eplication(dATP)m + (dCTP)n + (dGTP)y + (dGTP)z(dAMP + dCMP + dGMP + dGMP) m +n+ y+z + (m + n + y + z) PPi topoisomerasesingle strand binding proteinA C A G T G A T C A G AT G T C T C T5O P P P HPPPHOO H 35A533OOOHGhelicaseDNA polymerase DNA polymerase RNaseprimaseprimaseligasesingle strand binding

12、proteindGTP/dTTP/dCTP/dTTPeukaryotepolymeras *polymeras polymeras polymeras *polymeras prokaryotepolymeras Ipolymeras IIpolymeras III* veriest replicase substitute major action proof read, repair, filling major action in mitochondria DNA polymerases proof read, repair, filling substitute DNA polymer

13、ase III of E. coli DNA polymerase I of E. coliAGI1 109kD, 18 Helix 2 A-F: 323 aa, small fragment, 53 exonuclease activity3 G-R/c-terminal: 604 aa, Klenow fragment , 3 5 exonuclease activity, 5 3 polymerase activity, about 20 nucleotide.50 aaHOECDBRNPLMQKJNCFGD+ 35+PPi 35DNA polymerases function-153

14、polymerase activity AG53 Polymerase I Polymerase II Polymerase I Polymerase II Polymerase III Proof reading cut the fragment of primer andmutated fragments *DNA polymerases function-2 exonuclease activityCG53GTCAA53G53(1) The strict base complementaryC53T53high fidelity of DNA replicationThe conform

15、ation of DNA polymerase III is changeable,as its affinity to nucleotide acids (2) the character of DNA polymerase selecting base AACGT53ATGCATGCATGCATGCATGCGCATGCATGCTTT (3) The proof reading function of DNA polymerase A53TA53AA53CA53Gproofreading DNA helicase DNA topoisomerase DNA single strand bin

16、ding protein helicaseDna A, Dna B (rep), Dna CDna X Dna B Dna C Dna ADna T Dna C Dna T 1 2 3 4 5 6 7 8 9 10 11 helicaseDNA polimerase positive superhelix 1 2 3 4 5 6 7 8 9 10 11 negative superhelix1 2 3 4 5 6 7 8 9 10 11 topoisomerase normal helixDNA topoisomeraseuntwistingtopoisomerase I:breaks sin

17、gle strand DNA, not requires ATPtopoisomerase II:breaks double strands DNA, requires ATP cutligatesingle stranded DNA-binding protein, SSB (1) SSB is consist of 177 amino acid residues in E.coli(2) tetrapolymer(3) a SSB= 32 nucleotides(4) Cooperative bindingDna B SSB bank The role of SSBthe actions

18、of primer and primase3535DNA polymerase III53355353New DNA strand35Parental DNA templateprimaseSSBPrimase and Primosome The primase is a RNA polymerase, which is different with that in the transcription. The primase synthesizes the primers, which is used in DNA synthesis The primer is a fragment of

19、RNA The length of primer is 10-20bp approximately The helicases and other replicated factors have binded with DNA, then the primase have binds with them and formed a primosome at last. 3355 ATP ADP+Pi3355 ligaseOHPOH PThe compare of several enzymes, whichin formation of phosphodiester bondenzymespro

20、vides 3-OHprovides 5-Presults1 DNA primer or dNTP (dNMP) n+1 polymerase extending DNA strand 2 DNA no continued two single strands no continue ligase continue3 DNA to break and put in order two put in order to DNA topoisomerase DNA single strands superhelical structure4 primase extending primer NTP

21、primer A C A G T G A T C A G AT G T C T C T5O P P P HPPPHOO H 35A533OOOHGhelicaseDNA polymerase DNA polymerase RNaseprimaseprimaseligasesingle strand binding proteindGTP/dTTP/dCTP/dTTPPreparationFirst, you will read the summary in every chapters. Second, To the best of your as much as, to understand

22、 the tables and the figures in every chapters.Last, Find out the problems in them. The differences between the prokaryote and the eukaryote in DNA replication ?Section 3 Process of DNA replicationMG1G2S cell cycle initiation extension termination 1234Initiation of DNA replicationProkaryotic cell ( f

23、or instance E.coli)1 A fixed origin (ori) 2 Bidirectional replication Eukaryotic cella lot of origintheta12origin2 1Oorigin2 1OO212 1OO33DirectionDDDProkaryotic cellEukaryotic cellOriginHelicase, SSB .12lagging strandsleading strands1 13 17 29 32 4455 66 166 174 201 209 237 245TGTGGATTA-TTATACACA-TT

24、TGGATAA-TTATCCACAGATCTNTTTATTT-GATCTNTTNTATT-GATCTCTTATTACThe oriC of E coli.palindrome structureDnaADnaBDnaBDnaCDnaCDnaTDnaTThe extension of DNA replication rate of extension1 prokaryotic cell, 2500 bp/s。2 eukaryotic cell a lot of originpolymerase III holoenzymeprimerleading strandlagging strandpol

25、ymerase III subunit555333topoisomerasesingle strand binding proteinthe termination of DNA replication the terminate point of E.coli and SV40 virusoriginterminate point E.coliSV 40 82%32%0%50%Ligase 3535Pol IOHP3535RNaseOHP3535OHP3535Parental DNA template strandRNA primerNew DNA strandthe role of sev

26、eral enzyme in termination of DNA replication degradationligationfillingOoriginOOOO33DDDDDdirectionleading strandlagging strandOkazaki fragmentsThe 100-2000 bp Okazaki fragments was formed in DNA lagging strand in DNA replication . Rolling circle replication 13 OH535810235945367telomere and telomera

27、se of eukaryotesstructure of telomereTTAGGGTTAGGGTTAGGGTTAGGGsequence of telomereThe end of linear DNA in replication535CCCUAAGGGATTGGGATTstructure of telomeraseRNARTaseTP15335 CCCUAAGGGATTGGGATTmechanism of telomerase5335 CCCUAAGGGATTGGGATT5335 CCCUAA TTGGGATT5335 CCCUAA TTGGGATT5335 CCCUAA TTGGGAT

28、T53123435 CCCUAA TTGGGATT5335 CCCUAA TTGGGATT5353553 TTGGGATT3553 TTGGGATT3553 TTGGGATT TTGGGATT3535123Section 4 Reverse transcription and others single RNA strand RNA-DNA double strandssingle DNA strandDNA double strandsRTaseRNase HRTaseRNA355353RTase5353RNase H53integration synthesize cDNA from mR

29、NAAAAAAA53mRNAAAAAAA35AAAAAATTT.TTT5353primer: oligo dTreverse transcriptaseTTT.TTT53basic hydrolysisTTT.TTT53?TTT.TTT53AAAAAADNA polymerase ITTT.TTT53AAAAAA35S1 nucleasesignificance of reverse transcription ?Section 5DNA damage and repairmutationsThey are heritable permanentchanges in the base sequ

30、ence of DNA.DNA damagethe effect of mutation3 Lethal 4 Only lost some function 1 Useful 2 There may be some effect to the genotype, and no or not detectable effect to the phenotype cause of mutation1 spontaneous mutation mechanism? frequency 1/109 2 physical 3 chemical 4 biologicalInduced mutationmu

31、tational types 1 mismatch / point mutation 2 deletion 3 insertion 4 rearrangement / inversion point mutation1) transition2) transversion transitionA given pyrimidine is changed tothe other pyrimidine, or a givenpurine is changed to the other purine. transitionC TT CG AA GC GC G T 1C A T T AC A C 2C

32、T C GCCC A 3C T A A TC T G 4CC G ATG CCC CTA GGT CTC CTG TGG CTG GGC CTA GCC CTG TTG GGG GCT CTG CAT GCC CAG GCC CAG GAC TCC ACC TCA GAC CTG ATC CCA GCC CCA CCT CTG AGC AAG GTC CCT CTG CAG CAG AAC TTC CAG GAC AAC CAA TTC CAG GGG AAG TGG TAT GTG GTA GGC CTG GCA GGG AAT GCA ATT CTC AGA GAA GAC AAA GAC

33、 CCG CAA AAG ATG TAT GCC ACC G*TC TAT GAG CTG AAA GAA GAC AAG AGC TAC AAT GTC ACC TCC GTC CTG TTT AGG AAA AAG AAG TGT GAC TAC TGG ATC AGG ACT TTT GTT CCA GGT TGC CAG CCC GGC GAG TTC ACG CTG GGC AAC ATT AAG AGT TAC CCT GGA TTA ACG AGT TAC CTC GTC CGA GTG GTG AGC ACC AAC TAC AAC CAG CAT GCT ATG GTG TT

34、C TTC AAG AAA GTT TCT CAA AAC AGG GAG TAC TTC AAG ATC ACC CTC TAC GGG AGA ACC AAG GAG CTG ACT TCG GAA CTA AAG GAG AAC TTC ATC CGC TTC TCC AAA TCT CTG GGC CTC CCT GAA AAC CAC ATC GTC TTC CCT GTC CCA ATC GAC CAG TGT ATC GAC GGC TGASequence of NGAL gene in SHEEC cell223 AG, 75 IV transvertionTransversi

35、ons are changes from a pyrimidine to either of the two purines, and or the changes of a purine intoeither of the two pyrimidines.C GC AT GT Atransversion CGCG G 1CC G CGCG A 2CT A TACA G 3CT A TACA A 4CC G A CA TG CG TATCT C 5CG C ATCT T 6CT T GCCC C 7CG C GCCC T 8CA T HbA HbS genepeptidegenepeptide

36、GluValCTCCACGAGGTG17TA, 6GluValpoint mutation C A C G T A T GA CA TG CG T transversions transitions C TT CA GG Adeleted mutationinserted mutationinversed mutation55335533OHOHOHOH PPPP5533OHPP5533OHOHOHPPrearrangement 1 2 Hb genes family in 11 chromosome 1 2 1 2 1 2 1 2 Hb anti-LeporeHb Leporetwo gen

37、e types of Med anaemiarepair of DNA damaged1 photoreactivation repair2 excision repair3 recombination repair4 SOS repair Photoreactivation repairOOOOPHNNRHNNRCH3CH3UVphotolyase300-600nmOOORPHNNRCH3OHNCH3N excision repairaffectfactorspecificendo-nucleases35ligase5353pol 1Uvr AUvr BUvr C recombination

38、 repairpol IligaseRecASOS repair The DNA is damaged gravely and extensively. The replication of DNA had been stoped. The SOS repair is a emergency step. The SOS repair is a complicated network of regulation. The rate of DNA mutation is still very high. The SOS repair is called error prone repair.練習題

39、練習題DNA復制復制1、DNA復制時，下列哪一種酶是不需要的？復制時，下列哪一種酶是不需要的？ A DNA指導下的指導下的DNA聚合酶聚合酶 B DNA連接酶連接酶 C 拓撲異構(gòu)酶拓撲異構(gòu)酶 D 解鏈酶解鏈酶 E 限制性內(nèi)切酶限制性內(nèi)切酶2、合成、合成DNA的原料是的原料是 A dNMA B dNDP C dNTP D NTP E NMP3、真核細胞進行、真核細胞進行DNA復制的部位是復制的部位是 A 細胞膜細胞膜 B 細胞漿細胞漿 C 細胞核細胞核 D 核蛋白體核蛋白體 E 微粒體微粒體 原核細胞進行原核細胞進行DNA復制的部位是其擬核區(qū)復制的部位是其擬核區(qū) A 由由DNA為模板合成的為模板

40、合成的DNA片段片段 B 由由DNA為模板合成的為模板合成的RNA片段片段 C 由由RNA為模板合成的為模板合成的DNA片段片段 D 由由RNA為模板合成的為模板合成的RNA片段片段 4、DNA復制中的引物是復制中的引物是 A 5-TCTA-3 B 5-ATCT-3 C 5-UCUA-3 D 5-GCGA-3 E 3-TCTA-5 5、DNA復制時，模板序列復制時，模板序列 5-TAGA-3 將合成哪種互補序列？將合成哪種互補序列？ 6、關(guān)于、關(guān)于DNA復制中復制中DNA聚合酶的錯誤說法是聚合酶的錯誤說法是 A 底物是底物是dNTP B 必須有必須有DNA模板模板 C 合成方向是合成方向是5 3 D 需要需要Mg2+參與參與 E 需要需要ATP參與參與7、下列關(guān)于大腸桿菌、下列關(guān)于大腸桿菌DNA聚合酶聚合酶 的敘述哪一項是正確的的敘述哪一項是正確的